In order to adapt to teaching reform of Medical Immunology and create a"golden course",through teaching prac-tice,our research group proposes a"three-dimensional curriculum teaching system",which organically combines teaching concepts with teaching design and process,with construction of various resources(material libraries),and finally,multi-level teaching evalua-tion is completed to achieve overall unity of teaching.

In order to adapt to teaching reform of Medical Immunology and create a"golden course",through teaching prac-tice,our research group proposes a"three-dimensional curriculum teaching system",which organically combines teaching concepts with teaching design and process,with construction of various resources(material libraries),and finally,multi-level teaching evalua-tion is completed to achieve overall unity of teaching.

Objective Corruption is the most common cadaver phenomenon in forensic practice and an important basis for inferring time of death(PMI),but the definition of corruption degree and the construction of model inference models have always been difficult in the practice of forensic science.Methods In this study,the late postmortem phenomena were observed.Meanwhile,the microbial flora structure of gut and gravesoil and the nature of gravesoil were detected,for analyzing the changes before and after the key moment of abdominal rupture which naturally happened during the cadaver decay.Results The results found that from the macroscopic and microscopic levels,there were significant differences in cadaver decay,including microbial flora structure and gravesoil properties before and after the key moment of the natural abdominal rupture during cadaver decay.The phenomena are highly observable and can be accurately judged by forensic examinations,as well as related means in the field of biology and physiochemistry.In this study,this critical event was called Rupture Point.Conclusion The Rupture Point can be used as an important node for the assessment of cadaver decay degree in the practice of forensic medicine.It can be utilized for a cut-off point as well when constructing PMI inference models based on microbial flora structure changes.The accuracy of PMI inference models can be improved when the models were constructed in segments.

The DNA characterization of Cannabis sativa L.has been one of the key directions of anti-drug research at home and abroad.Previous research mainly focused on the identification of cannabis-species and gender differentiation,and have constructed a number of corresponding composite amplification systems.With the rapid development of high-throughput sequencing technology,the whole genome of C.sativa and the sequences of key enzyme genes for its major physicochemical components have been sequenced successively,and intra-species differentiation studies of C.sativa based on specific molecular markers have gradually emerged.However,due to the high variability of cannabis subspecies-and variety-specific molecular markers,relevant foreign studies failed to provide ideal molecular marker support for the identification of intra-specific distinctions of Cannabis sativa in China.Based on this,this paper comprehensively analyzes the current situation and shortcomings of domestic and international research on intra-specific differentiation of C.sativa,and combines the previous research results of this group to elaborate on how to use high-throughput sequencing technology to solve the problem of the lack of intra-specific molecular markers of C.sativa in China.

OBJECTIVETo investigate the effect of laparoscopic sleeve gastrectomy(LSG) on sex hormone in male patients with severe obesity.

METHODSRetrospective analysis was performed in 31 male patient with severe obese [body mass index(BMI) ≥28 kg/m, obesity group] who underwent LSG in Shanghai Tenth People's Hospital of Tongji University from December 2012 to May 2016. The anthropometric parameters(weight, BMI, waist circumference, hip circumference, waist-hip ratio, body fat percentage), glucose metabolic indices [fasting plasma glucose(FPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index(HOMA-IR)], and sex hormone parameters [estradiol(E2), total testosterone (TT), follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] were collected preoperatively and 1, 3, 6 months postoperatively. In addition, 31 healthy male volunteers with normal BMI were consecutively recruited in this study as control group. The above-mentioned parameters were also determined in control group. Changes of these variables before and after surgery were analyzed. Pearson method was used to analyze the correlation of TT with anthropometric parameters and glucose metabolic indices before and after surgery.

RESULTSThe average age of patients in obesity and control group was (32.9±9.7) (18 to 56) years and (30.7±8.9) (18 to 49) years. Compared to the control group, obesity group had significantly higher anthropometric parameters and glucose metabolic indices before surgery (all P<0.05). In obesity group, the anthropometric and glucose metabolic indices significantly decreased at 1 to 6 months after surgery compared to those before surgery (all P<0.05). At 1 month after surgery, the anthropometric parameters and glucose metabolic indices in obesity group were significantly higher than those in control group (all P<0.05). At 3, and 6 months after surgery, there were no significant differences in glucose metabolic indices between obesity and control group (all P>0.05), while the anthropometric parameters in obesity group were still significantly higher than those in control group(all P<0.05). The sex hormone parameters in control and obesity group before surgery were as follows: E2: (100.2±23.5) pmol/L and (129.2±81.9) pmol/L; TT: (18.0±4.9) nmol/L and (8.4±4.5) nmol/L; FSH: (4.5±3.1) IU/L and (4.3±2.5) IU/L; LH: (4.4±1.7) IU/L and (5.3±2.6) IU/L. Compared to control group, the TT level of obese patients before surgery significantly decreased(P=0.000), while no significant differences were observed in the levels of E2, FSH, and LH(all P>0.05). The TT levels were significantly increased at 1, 3, 6 months after surgery[(13.1±7.0), (13.6±5.7), (21.0±19.3) nmol/L, respectively, all P<0.05] and the E2 level was significantly decreased at 6 months after surgery [(91.4±44.9) pmol/L, P<0.05], while no significant differences were observed at 1 and 3 months after surgery (all P>0.05). Furthermore, the FSH and LH levels did not exhibit significant change at 1, 3, and 6 months after surgery compared to those before surgery (all P>0.05). At 1 month after surgery, no significant correlations were examined in the change value of TT levels (▹TT) with the changes of BMI(▹BMI), FPG(▹FPG), FINS(▹FINS), HOMA-IR(▹HOMA-IR), and E2(▹E2) (all P>0.05). At 3 months after surgery, ▹TT was negatively correlated with ▹BMI (r=-0.441, P=0.015), ▹FINS (r=-0.375, P=0.041), and ▹HOMA-IR(r=-0.397, P=0.030), but not correlated with ▹FPG and ▹E2 (all P>0.05). At 6 months after surgery, ▹TT was negatively correlated with ▹BMI(r=-0.510, P=0.018) and ▹HOMA-IR (r=-0.435, P=0.049), but not correlated with ▹FPG, ▹FINS and ▹E2 (all P>0.05).

CONCLUSIONSMale severe obese patients are accompanied with abnormal sex hormone levels. LSG has a significant effect on weight loss and blood glucose improvement, and may ameliorate the sex hormone unbalance by improving the insulin resistance in men with severe obesity.

Adult ; Bariatric Surgery ; Blood Glucose ; physiology ; Body Mass Index ; Body Weights and Measures ; China ; Estradiol ; blood ; physiology ; Fasting ; blood ; Follicle Stimulating Hormone ; blood ; physiology ; Follow-Up Studies ; Gastrectomy ; Glycated Hemoglobin A ; physiology ; Humans ; Insulin ; blood ; physiology ; Insulin Resistance ; physiology ; Luteinizing Hormone ; blood ; physiology ; Male ; Obesity, Morbid ; surgery ; Retrospective Studies ; Testosterone ; blood ; physiology ; Treatment Outcome ; Weight Loss ; physiology

Objective To construct a rapid genetic DNA extraction method, with nano magnetic beads, self-designed reagents system and extracting process. Method Part I: DNA extraction from old blood cotton swab sample with self-designed DNA extraction kit, then quantiifed by UV spectrophotometer. The method was further optimized on the preliminary results. Part II: All kinds of difficult DNA sample were tested with optimized kit, to detect the applicability of the kit. Result By improving the experimental condition, the extraction effects of different DNA sample is good, meanwhile, the extraction cost is relatively low.

Objective To report our first clinical experience with a novel modified culotte technique for the treatment of true coronary bifurcation lesions. Methods The novel modified culotte technique (the mono-ring culotte) stenting was done in which the side branch (SB) stent was deployed firstly followed by ex vivo wiring of a most proximal cell of SB stent with the hard end of main branch (MB) wire. Secondly, the MB stent was deployed through the most proximal cell of SB stent. The procedure was ended with kissing balloon dilation. From June 2014 to March 2015, 15 patients with true coronary bifurcation lesion were treated with mono-ring culotte stenting in our center. Results The procedures were successful in all cases without procedural complication and in-hospital major adverse cardiovascular events. The procedural time was (34. 3 ± 9. 6) min, fluoroscopic time was (18. 1 ± 3. 8) min, and contrast volume was (112. 0 ± 24. 5) ml, respectively. Post-procedurally, the residual stenosis of the main and the side branch were (10. 0 ± 2. 5)% and (10. 2 ± 5. 3)% , respectively. Conclusions The mono-ring culotte stenting is safe and feasible for treatment of true coronary bifurcation lesions, and may be superior to the conventional culotte stenting.

Gene cloning is one of the most important and widely used technologies in molecular biology research. Generally, DNA fragment is cut with restriction enzyme, and then the product is ligated to a linearized vector with complementary sticky end or blunt end by DNA-ligase. This traditional DNA cloning method requires compatible enzyme recognition sites existing in both PCR fragment and targeted vector. Several ligase-free methods have been established to avoid the using of restriction enzyme. However, those methods are time-consuming, labor-intensive and expensive. To overcome these shortcomings, we developed an Exonuclease III based DNA cloning method that takes only 30 minutes with high cloning efficiency and significant economic advantage. Therefore, this method is suitable for large-scale gene cloning.
Cloning, Molecular ; methods ; DNA ; chemistry ; DNA Restriction Enzymes ; Exodeoxyribonucleases ; chemistry ; Genetic Vectors ; Polymerase Chain Reaction

Most current research in the field of adventitious shoot formation is focused on the regulatory function of a single gene. However, a systematic transcriptomic analysis of the early adventitious shoot formation is still lacking. Here, we analyzed the transcriptome profiling of the early adventitious shoot formation in Arabidopsis by RNA-seq high throughput sequencing technology, and identified 2 457 differentially expressed genes. Detailed categorization revealed that these genes were mainly involved in hormone homeostasis or signal transduction, callus and lateral root formation, shoot apical meristem development and photosynthesis. Further pathway enrichment analysis showed that genes involved in phenylalanine metabolism and phenylpropanoid biosynthesis were significantly enriched. Moreover, exogenous phenylalanine could repress adventitious shoot formation, indicating that phenylalanine metabolism and phenylpropanoid biosynthesis might be important for adventitious shoot formation.
Arabidopsis ; genetics ; Gene Expression Profiling ; methods ; Gene Expression Regulation ; Genes, Plant ; Phenylalanine ; pharmacology ; Plant Shoots ; genetics ; growth & development

Objective To investigate the expression of tumor suppressor gene p16~(INK4A) in mouse endometrium during the estrus cycle and early pregnancy and its possible role in blastocyst implantation. Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of p16~(INK4A) mRNA,immunohistochemistry and Western blotting were applied to detect p16~(INK4A) protein in mouse endometrium tissues during the estrus cycle and early pregnancy. Results The intensity of p16~(INK4A) mRNA expression in mouse early pregnancy was higher than that in the estrus cycle.Compared with the other 3 stages, the level of p16~(INK4A) mRNA expression at estrus was obviously higher. During the early pregnancy, the level of p16~(INK4A) mRNA expression increased steadily from day 2 to day 5,reaching the maximal level on day 5,then decreasing. Both immunohistochemical and Western blotting analysis showed the same results in expression patterns of p16~(INK4A) protein for mouse endometrium tissues as those results of RT-PCR.Conclusion p16~(INK4A) is involved in the embryos penetrating into the endometrial barrier.